Easy detection of green fluorescent protein multicopy transformants in Penicillium griseoroseum

Show simple item record Lopes, Francis J.F. Araújo, Elza F. de Queiroz, Marisa V. de 2017-10-24T09:32:26Z 2017-10-24T09:32:26Z 2004-12-21
dc.identifier.issn 16765680
dc.description Não possui URI pt-BR
dc.description.abstract Penicillium griseoroseum, a deuteromycete fungus producer of pectinolytic enzymes, was transformed with a gene encoding for green fluorescent protein (GFP). The selection of transformants was based on the homologous nitrate reductase gene (niaD). Protoplasts of a P. griseoroseum Nia mutant (PG63) were co-transformed with the plasmids pNPG1 and pAN52-1-GFP. The plasmid pNPG-1 carries the homologous niaD gene and pAN52-1-GFP carries the SGFP-TYG version of GFP. The highest transformation efficiency (102 transformants/µg of pNPG1) resulted from the utilization of equimolar amounts of transforming and co-transforming vectors. Analysis of pAN52-1-GFP insertions into the genomic DNA of the transformants revealed single and multiple copy integrations. The transformants possessing a single copy of the gfp gene showed a low level of fluorescence, whereas multicopy transformants displayed strong fluorescence under visualization with fluorescent light. The transformants showing high expression of the gfp gene had the normal mycelia pigmentation altered, displaying a bright green-yellowish color, visible with the naked eye on the plates, without the aid of any kind of fluorescent light or special filter set. en
dc.format pdf pt-BR
dc.language.iso eng pt-BR
dc.publisher Genetics and Molecular Research pt-BR
dc.relation.ispartofseries 3 (4), p.449-455, December 2004 pt-BR
dc.rights Open Access pt-BR
dc.subject Penicillium griseoroseum pt-BR
dc.subject Transformation pt-BR
dc.subject Protoplasts pt-BR
dc.subject GFP pt-BR
dc.subject Pectinases pt-BR
dc.subject Mutant pt-BR
dc.title Easy detection of green fluorescent protein multicopy transformants in Penicillium griseoroseum en
dc.type Artigo pt-BR

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    Artigos Técnico-científicos na área de Microbiologia

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